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Safe and easy in vitro evaluation of tmRNA-SmpB-mediated -translation from ESKAPE pathogenic bacteria

RNA. 2021-08; 
Marion Thépaut, Rodrigo Campos-Silva, Eva Renard, Frédérique Barloy-Hubler, Eric Ennifar, Daniel Boujard, Reynald Gillet
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Codon Optimization … For each ESKAPE SmpB, GenScript synthesized the sequences with codon optimization for E. coli, cloning them into the pET22b(+) vector between the NdeI and XhoI restriction sites to add a 6His histidine tag (Supplemental Table 2). The generated plasmids, … Get A Quote

摘要

In bacteria, -translation is the major quality control system for rescuing stalled ribosomes. It is mediated by tmRNA, a hybrid RNA with properties of both a tRNA and a mRNA, and the small protein SmpB. Because -translation is absent in eukaryotes but necessary for bacterial fitness or survival, it is a promising target for the development of novel antibiotics. To facilitate screening of chemical libraries, various reliable in vitro and in vivo systems have been created for assessing -translational activity. However, the aim of the current work was to permit the safe and easy in vitro evaluation of -translation from pathogenic bacteria, which are obviously the ones we should be targeting. Based on green fluores... More

关键词

ESKAPE, HTS, antibiotics, ribosome, tmRNA, trans-translation